期刊论文详细信息
Molecules
Thermal Shift Assay as a Tool to Evaluate the Release of Breakdown Peptides from Cowpea β-Vignin during Seed Germination
Giuditta C. Heinzl1  Alessandro L. Heinzl1  Matias Pasquali1  Stefano De Benedetti1  Daniela Lupi1  Camilla Leogrande1  Francesco Castagna1  Alessio Scarafoni1 
[1]Department of Food, Environmental and Nutritional Sciences, University of Milan, 20133 Milan, Italy
关键词: seed storage proteins;    thermal stability;    germination;    proteolysis;    bioactivity;    Vigna unguiculata;   
DOI  :  10.3390/molecules27010277
来源: DOAJ
【 摘 要 】
The present work aimed to characterize the molecular relationships between structure and function of the seed storage protein β-vignin, the vicilin storage protein of cowpea (Vigna unguiculata, l. Walp) seeds. The molecular characterization of β-vignin was carried out firstly by assessing its thermal stability, under different conditions of pH and ionic strength, by thermal shift assay (TSA) using SYPRO Orange fluorescent dye. Secondly, its aggregation propensity was evaluated using a combination of chromatographic and electrophoretic techniques. Two forms of β-vignin were considered: the native form purified from mature quiescent seeds, and a stable breakdown intermediate of 27 kDa produced while seeds germinate. TSA is a useful tool for determining and following over time the structural changes that occur to the protein during germination. The main result was the molecular characterization of the 27 kDa intermediate breakdown polypeptide, which, to the best of our knowledge, has never been described before. β-vignin seems to retain its trimeric conformation despite the evident degradation of its polypeptides.
【 授权许可】

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